Parasporins are Cry toxins from Bacillus thuringiensis that are active against tumor cells. This work shows that parasporin PS1Aa2 (Cry31Aa2) forms pores in artificial membranes like many Cry toxins. These pores have several levels of conductance; the most frequently seen in 150 mM KCl solutions were of 11, 16 and 21 pS. Our
microspectrofluorometric results with the Fura-2 probe showed that the presence of PS1Aa2 can induce changes in intracellular calcium levels, most often in the form of calcium oscillations and sometimes as sustained increases. Such responses were observed in the presence and absence of extracellular calcium, with the tumor cell lines HeLa and HepG2, and with the non-tumorous cell line HEK 293. Calcium oscillations have not been described previously for Cry toxins even though some studies have shown that calcium appears to be involved in their mode of action. Our experiments required the use of much higher concentrations of toxin than suggested from the published cytotoxicity results. Despite the presence of fragments previously identified as active, its low efficacy appears to be related to the presence of DNA in the preparations causing the protein to precipitate. Future work on this toxin would therefore be greatly facilitated by an improvement in its method of preparation.